TGF-β与BMP复合材料的骨修复作用.doc

TGF-与BMP复合材料的骨修复作用TGF-与BMP复合材料的骨修复作用.txt爱情是艺术，结婚是技术，离婚是算术。这年头女孩们都在争做小“腰”精，谁还稀罕小“腹”婆呀？高职不如高薪，高薪不如高寿，高寿不如高兴。TGF-与BMP复合材料的骨修复作用 2010-02-25 孙玉鹏张皖清马福成陈万禄 【摘要】目的探讨转化生长因子(TGF-)与骨形成蛋白(BMP)复合后对节段性骨缺损的修复作用,以及在骨修复过程中TGF-和BMP之间的相互作用。方法采用以系列化学方法处理的小牛松质骨作为载体,将一定量的TGF-和/或BMP与之复合,形成TGF-/载体、BMP/载体和TGF-/BMP/载体复合物(其中含TGF- 120g、BMP12mg),分别修复1.5cm兔桡骨节段性骨缺损。术后4、8、12、16周取材,进行放射学和组织学观察。结果术后4周,植入TGF-/载体的缺损区内可见不均匀絮状放射阴影,外骨膜呈增生反应,骨折线模糊不清,缺损区尚未完全桥接;植入BMP/载体组新生骨痂明显,密度均匀,缺损区两断端骨折线隐约可见;在植入TGF-/BMP/载体的缺损区可见均匀的高密度阴影,桥接骨缺损的两端,骨折线模糊。术后第1216周3组缺损区完全性修复,其密度近正常骨,可见皮质骨和骨髓腔结构。16周时TGF-和TGF-/BMP组骨缺损区周围外骨痂少,而BMP组中仍可见较多尚未完全吸收的外骨痂。结论TGF-/BMP复合后早期骨修复作用优于单一的TGF-和BMP,在骨缺损修复过程中TGF-和BMP有良好的协同作用。 【关键词】转化生长因子骨形成蛋白骨缺损骨修复骨生成 Effect of Composite of Transforming Growth Factor and Bone Morphogenetic Protein on Repairing Bone DefectsSUN Yu-peng, ZHANG Wan-qing, MA Fu-cheng, et al. Center of Orthopaedics, 304th Hospital of PLA, Beijing 100037 【Abstract】AimTo evaluate the effect of transforming growth factor (TGF-) and bone morphogenetic protein (BMP) composite on the repairing of large segmental bone defects and to discuss the interaction between TGF- and BMP during bone repair. MethodsSegmental bone defects for 1.5cm were created in the mid-upper part of the radial shafts of 48 adult rabbits. The defects were filled with implants of TGF-/carrier, BMP/carrier, and TGF-/BMP/carrier (TGF- 120Ug, BMP 12mg) separately. The defects were examined radiographically and histologically at 4, 8, 12 and 16 weeks post-operation. ResultsIn the group with TGF-/BMP composite, the defects area was bridged with callus of uniform radiodensity at 4 weeks, and cortices of the cut ends were obscured by new bones. At 16 weeks post-operation, the defects were bridged by uniform new bones and the cut ends of cortex could not be seen in all groups. In the group with BMP/carrier, the defects were filled with more irregular woven bone callus than those in the other two groups. Defects implanted with TGF-/BMP in animals killed at 16 weeks showed histologically new lamellar and woven bone which was formed in continuity with the cut ends of the cortex. Medullar canal was recanalized and contained marrow elements with normal appearance. ConclusionThese data demonstrate not only the synergistic action between TGF-/BMP in the process of bone healing, but also a better effect of TGF-/BMP composite than single TGF- or BMP on bone repair, especially in the early stage of bone repair process. 【Key words】Transforming growth factor Bone morphogenetic proteinBone defectBone repairOsteogenesis 骨修复是一个极其复杂而有序的过程。近年的研究表明,生长因子在骨愈合过程中起重要作用。骨形成蛋白(BMP)是骨生成的启动因子,能诱导间充质细胞分化成软骨细胞和骨细胞,对骨愈合有明显促进作用。转化生长因子(TGF-)机体内含量最为丰富,在胚胎发育、创口愈合、基质形成、骨组织形成及改建过程中都起着重要作用。笔者以前的实验显示,TGF-刺激间充质细胞的增殖和分化,形成成骨细胞和软骨细胞,从而诱导新骨的生成,并且具有修复大块节段性骨缺损的能力1，2。本实验旨在探讨TGF-/BMP复合材料对骨缺损的修复作用及TGF-和BMP在骨修复中的相互作用。 材料与方法 1.复合材料的制备3:以新鲜牛血为原料,离心,分离出血小板。采用酸醇抽提方法,提取血小板蛋白,并先后经过2次Bio-Gel P60柱层析,分离纯化出TGF-1。本实验采用的BMP由已建立的方法从小牛皮质骨中提取;选择经化学处理的牛松质骨作为载体。将一定量的TGF-和/或BMP分别溶解后与载体在真空条件下进行充分混匀,形成3种复合物,即TGF-/载体、BMP/载体和TGF-/BMP/载体复合物。冻干,射线照射消毒。 2.动物实验:健康家兔48只,体重2.53.0kg,雌雄不分。随机分成3组,每组18只。在3%戊巴比妥钠静脉麻醉下,手术显露右桡骨中上段,连同骨膜一起切除,造成桡骨1.5cm节段性骨缺损。以等渗盐水清洗创口后,分别植入TGFC-/载体、BMP/载体和TGF-/BMP/载体复合材料60mg(其中分别含TGF- 120g、BMP 12mg)。逐层缝合伤口。术后不给抗生素,不加外固定。动物分笼饲养,常规饮食。 3.观察指标:(1)X线检查:分别在术后4、8、12和16周处死动物进行取材。在48kV、25mA、0.12s、75cm的投照条件下由专人作X线拍片及冲洗,观察各组动物骨缺损区的骨痂生长和桥接情况,并根据其愈合程度按Cook等6级评分标准4进行量化评估。(2)组织学观察:标本以10%甲醛固定后,混合脱钙液脱钙处理,石蜡包埋切片,HE染色,观察比较各组骨缺损愈合进程,依照Nilsson等骨愈合组织学评分标准5进行评分。 4.统计学处理:对X线及组织学评分结果作秩和检验处理(SPLM软件包,第四军医大学统计学教研室)。 结果 1.X线检查结果:术后4周,植入TGF-/载体骨缺损区内可见不均匀的放射阴影,外骨膜呈增生反应,骨折线模糊不清,缺损区尚未完全桥接;植入BMP/载体组新生骨痂较明显,密度均匀,缺损区骨折线隐约可见;植入TGF-/MBP/载体的缺损区可见均匀的高密度阴影桥接骨缺损的两端,骨折线模糊(图13)。第8周时,TGF-组骨缺损区阴影密度明显增高